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. 2014 Oct 24;6(15):13822–13834. doi: 10.18632/oncotarget.2635

Figure 4. Silencing the expression of ANGPTL2 reverts EMT in HPDE and HPNE /KRAS, /KRAS/HER2/p16p14shRNA expressing cell lines.

Figure 4

A, results of quantitative real-time PCR analysis of ANGPTL2 gene expression in HPDE and HPNE, /KRAS, /KRAS/HER2/p16p14shRNA cell lines expressing shRNA sequences to knock down the expression of ANGPTL2 or scramble sequence as control. Results are presented as the fold change in RNA expression between the gene of interest and β-actin. The mean values and SEM from 3 independent experiments conducted in quadruplicate are shown. ***, P < 0.001 by two-tailed unpaired Student's t tests. B, ELISA analysis of the secretion of ANGPTL2 in the conditional medium of HPDE and HPNE, /KRAS, /KRAS/HER2/p16p14shRNA cell lines expressing shRNA sequences to knock down the expression of ANGPTL2 or scramble sequence as control. Results are presented as percentages of the total protein extract from the respective secreting cells. The mean values and SEM from 3 independent experiments conducted in quadruplicate are shown. ***, P < 0.001 by two-tailed unpaired Student's t tests. C, results of quantitative real-time PCR analysis of CDH1 gene expression in HPDE, HPDE/KRAS and HPDE/KRAS/HER2/p16p14shRNA cell lines expressing shRNA sequences to knock down the expression of ANGPTL2 or scramble sequence as control. D, results of quantitative real-time PCR analysis of vimentin gene expression in HPNE, HPNE/KRAS and HPNE/KRAS/HER2/p16p14shRNA cell lines expressing shRNA sequences to knock down the expression of ANGPTL2 or scramble sequence as control. Results are presented as the fold change in RNA expression between the gene of interest and β-actin. The mean values and SEM from 3 independent experiments conducted in quadruplicate are shown. ***, P < 0.001 by two-tailed unpaired Student's t tests. E, Western blot analysis for the expression of E-cadherin and vimentin in HPDE and HPNE, /KRAS and /KRAS/HER2/p16p14shRNA cell lines expressing shRNA sequences to knock down the expression of ANGPTL2 or scramble sequence as control. The relative protein quantification was reported as ratio with γ-tubulin protein expression levels.